Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding

Michael E. Maguire; Alfred G. Gilman

doi:10.1016/0005-2744(74)90267-8

Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding

Michael E. Maguire, Alfred G. Gilman

Pharmacology

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

An assay for adenylate cyclase (EC 4.6.1.1) is described using adenylyl imidodiphosphate (AMP-PNP) as substrate. Methods are described for introducing the cyclase reaction mixture directly into a protein binding assay for the cyclic nucleotide without purification of cyclic AMP. Significant substrate depletion and regenerating systems are avoided by this method, and blank values are negligible. The assay is capable of reproducibly detecting adenylate cyclase activity with less than 5 μg of protein from rat cerebral cortex.

Original language	English (US)
Pages (from-to)	154-163
Number of pages	10
Journal	BBA - Enzymology
Volume	358
Issue number	1
DOIs	https://doi.org/10.1016/0005-2744(74)90267-8
State	Published - Jul 17 1974

ASJC Scopus subject areas

Medicine(all)

Access to Document

10.1016/0005-2744(74)90267-8

Cite this

@article{b38e0682ea27464b8df9c788d08d1fb4,

title = "Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding",

abstract = "An assay for adenylate cyclase (EC 4.6.1.1) is described using adenylyl imidodiphosphate (AMP-PNP) as substrate. Methods are described for introducing the cyclase reaction mixture directly into a protein binding assay for the cyclic nucleotide without purification of cyclic AMP. Significant substrate depletion and regenerating systems are avoided by this method, and blank values are negligible. The assay is capable of reproducibly detecting adenylate cyclase activity with less than 5 μg of protein from rat cerebral cortex.",

author = "Maguire, {Michael E.} and Gilman, {Alfred G.}",

note = "Funding Information: work. These studies were supported by U.S. Public Health Service grants NS 10193 and GRS 5S01RR05431.",

year = "1974",

month = jul,

day = "17",

doi = "10.1016/0005-2744(74)90267-8",

language = "English (US)",

volume = "358",

pages = "154--163",

journal = "BBA - Enzymology",

issn = "0005-2744",

publisher = "Elsevier BV",

number = "1",

}

TY - JOUR

T1 - Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding

AU - Maguire, Michael E.

AU - Gilman, Alfred G.

N1 - Funding Information: work. These studies were supported by U.S. Public Health Service grants NS 10193 and GRS 5S01RR05431.

PY - 1974/7/17

Y1 - 1974/7/17

N2 - An assay for adenylate cyclase (EC 4.6.1.1) is described using adenylyl imidodiphosphate (AMP-PNP) as substrate. Methods are described for introducing the cyclase reaction mixture directly into a protein binding assay for the cyclic nucleotide without purification of cyclic AMP. Significant substrate depletion and regenerating systems are avoided by this method, and blank values are negligible. The assay is capable of reproducibly detecting adenylate cyclase activity with less than 5 μg of protein from rat cerebral cortex.

AB - An assay for adenylate cyclase (EC 4.6.1.1) is described using adenylyl imidodiphosphate (AMP-PNP) as substrate. Methods are described for introducing the cyclase reaction mixture directly into a protein binding assay for the cyclic nucleotide without purification of cyclic AMP. Significant substrate depletion and regenerating systems are avoided by this method, and blank values are negligible. The assay is capable of reproducibly detecting adenylate cyclase activity with less than 5 μg of protein from rat cerebral cortex.

UR - http://www.scopus.com/inward/record.url?scp=0016169228&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0016169228&partnerID=8YFLogxK

U2 - 10.1016/0005-2744(74)90267-8

DO - 10.1016/0005-2744(74)90267-8

M3 - Article

C2 - 4368377

AN - SCOPUS:0016169228

SN - 0005-2744

VL - 358

SP - 154

EP - 163

JO - BBA - Enzymology

JF - BBA - Enzymology

IS - 1

ER -

Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this